清酱肉多肽的抗氧化活性和氨基酸分析
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摘要
以清酱肉为研究对象,提取并通过超滤分离得到清酱肉多肽(分子质量<10 kDa),测定不同质量浓度多肽液对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基、羟自由基(·OH)的清除效果、Fe2+螯合能力和还原能力,分析清酱肉多肽的氨基酸组成。结果表明:当清酱肉多肽液的质量浓度为5 mg/mL时,其对·OH和DPPH自由基的清除率分别可达51.49%和57.50%,与相同质量浓度的谷胱甘肽(glutathione,GSH)相比差异显著(P<0.05);清酱肉多肽的Fe2+螯合能力随其质量浓度的增加而显著提高,当质量浓度为5 mg/mL时,其对Fe2+的螯合率可达50.97%;清酱肉多肽还具有一定的还原性。此外,清酱肉多肽液的氨基酸组成较为丰富,其中必需氨基酸含量为29.14%;与抗氧化活性相关的碱性、酸性及疏水性氨基酸的总量达到87.75%。综上所述,清酱肉多肽液具有一定的抗氧化能力及营养价值。
In this experiment, peptides(molecular mass < 10 kDa) were extracted from pickled sauced meat and separated by ultrafiltration. Antioxidant activities of the peptides were evaluated by determining scavenging activities against1,1-diphenyl-2-picrylhydrazyl(DPPH) and hydroxyl radicals, Fe2+ chelating capacity and reducing power. Furthermore, the amino acid composition was analyzed. The results showed that the percentage scavenging of hydroxyl and DPPH radicals by the peptides at 5.0 mg/mL was 51.49% and 57.50%, respectively, highly signi?cantly different from that by glutathione(GSH) at the same concentration(P < 0.05). The Fe2+ chelating capacity of the peptides was signi?cantly concentrationdependently enhanced, and at a concentration of 5 mg/mL, the percentage chelating of Fe2+ was 50.97%. Also the peptides showed reducing power. In addition, the amino acid composition of the peptides was abundant, with essential amino acids accounting for 29.14% of the total amino acids. The content of antioxidant amino acids including acidic amino acid, basic amino acid and hydrophobic amino acid was as high as 87.75%. In summary, the peptides extracted from pickled sauced meat has antioxidant activity and nutritional value.
In this experiment, peptides(molecular mass < 10 kDa) were extracted from pickled sauced meat and separated by ultrafiltration. Antioxidant activities of the peptides were evaluated by determining scavenging activities against1,1-diphenyl-2-picrylhydrazyl(DPPH) and hydroxyl radicals, Fe2+ chelating capacity and reducing power. Furthermore, the amino acid composition was analyzed. The results showed that the percentage scavenging of hydroxyl and DPPH radicals by the peptides at 5.0 mg/mL was 51.49% and 57.50%, respectively, highly signi?cantly different from that by glutathione(GSH) at the same concentration(P < 0.05). The Fe2+ chelating capacity of the peptides was signi?cantly concentrationdependently enhanced, and at a concentration of 5 mg/mL, the percentage chelating of Fe2+ was 50.97%. Also the peptides showed reducing power. In addition, the amino acid composition of the peptides was abundant, with essential amino acids accounting for 29.14% of the total amino acids. The content of antioxidant amino acids including acidic amino acid, basic amino acid and hydrophobic amino acid was as high as 87.75%. In summary, the peptides extracted from pickled sauced meat has antioxidant activity and nutritional value.
引文
[1]JOHANSENJS,HARRISAK,RYCHLYDJ,etal.Oxidative stressandtheuseofantioxidantsindiabetes:linkingbasicscience to clinical practice[J]. Cardiovascular Diabetology, 2005, 4(1):5-16.DOI:10.1186/1475-2840-4-5.
[2]REICHMANND,VOTHW,JAKOBU.Maintainingahealthy proteome during oxidative stress[J]. Molecular Cell, 2018, 69(2):203-213.DOI:10.1016/j.molcel.2017.12.021.
[3]李兴太,张春英,仲伟利,等.活性氧的生成与健康和疾病关系研究进展[J].食品科学, 2016, 37(13):257-270. DOI:10.7506/spkx1002-6630-201613046.
[4]GüL?IN?. Antioxidant activity of food constituents:an overview[J].ArchivesofToxicology,2012,86(3):345-391.DOI:10.1016/j.peptides.2010.06.020.
[5]SARMADIBH,ISMAILA.Antioxidativepeptidesfrom foodproteins:areview[J].Peptides,2010,31(10):1949-1956.DOI:10.1016/j.peptides.2010.06.020.
[6]ZHU Chaozhi, ZHANG Wangang, KANG Zhuangli, et al. Stability of an antioxidant peptide extracted from Jinhua ham[J]. Meat Science,2014, 96(2):783-789. DOI:10.1016/j.meatsci.2013.09.004.
[7]XINGLujuan,HUYaya,HUHongyan,etal.Purificationand identificationofantioxidativepeptidesfromdry-curedXuanwei ham[J].FoodChemistry,2016,194:951-958.DOI:10.1016/j.foodchem.2015.08.101.
[8]WANG Lusha, HUANG Jichao, CHEN Yulian, et al. Identi?cation and characterization of antioxidant peptides from enzymatic hydrolysates of duck meat[J]. Journal of Agricultural and Food Chemistry, 2015,63(13):3437-3444. DOI:10.1021/jf506120w.
[9]FENG Li, QIAO Yan, ZOU Yufeng, et al. Effect of?avourzyme on proteolysis,antioxidantcapacityandsensoryattributesofChinese sausage[J].MeatScience,2014,98(1):34-40.DOI:10.1016/j.meatsci.2014.04.001.
[10]ZHU Chaozhi, ZHANG Wangang, ZHOU Guanghong, et al. Isolation andidentificationofantioxidantpeptidesfromJinhuaham[J].Journal of Agricultural and Food Chemistry, 2013, 61(6):1265-1271.DOI:10.1021/jf3044764.
[11]孔令明,李芳,陶永霞,等.多肽的功能活性与研究进展[J].中国食品添加剂,2 0 0 9,6 7(3):7 1-7 3.D O I:1 0. 3 9 6 9/j.issn.1006-2513.2009.03.015.
[12]罗雨婷,吴宝森,谷大海,等.不同加工年份诺邓火腿粗肽的特征与抗氧化活性[J].肉类研究, 2017, 31(9):32-37. DOI:10.7506/rlyj1001-8123-201709006.
[13]赵改名,周光宏,柳艳霞,等.肌肉非蛋白氮和游离氨基酸在金华火腿加工过程中的变化[J].食品科学, 2006, 27(2):33-37.DOI:10.3321/j.issn:1002-6630.2006.02.001.
[14]MORAL,BOLUMART,HERESA,etal.Effectofcookingand simulatedgastrointestinaldigestionontheactivityofgenerated bioactivepeptidesinagedbeefmeat[J].FoodandFunction,2017,8(12):4347-4355. DOI:10.1039/C7FO01148B.
[15]GALLEGOM,MORAL,HAYESM,etal.Effectofcooking andinvitrodigestionontheantioxidantactivityofdry-curedham by-products[J].FoodResearchInternational,2017,97:296-306.DOI:10.1016/j.foodres.2017.04.027.
[16]SUN Weizheng, ZHAO Haifeng, ZHAO Qiangzhong, et al. Structural characteristics of peptides extracted from Cantonese sausage during dryingandtheirantioxidantactivities[J].InnovativeFoodScience andEmergingTechnologies,2009,10(4):558-563.DOI:10.1016/j.ifset.2009.07.006.
[17]陈好春,周兴虎,康大成,等.风味蛋白酶对腌腊鸡腿蛋白质降解、抗氧化能力和感官品质的影响[J].食品与发酵工业, 2016(7):128-134. DOI:10.13995/j.cnki.11-1802/ts.201607022.
[18]杨凯,李迎楠,李享,等.基于计算机视觉技术分析猪肉对清酱肉挥发性风味成分的影响[J].肉类研究, 2018, 32(4):39-43.DOI:10.7506/rlyj1001-8123-201804007.
[19]祝超智,张万刚,徐幸莲,等.金华火腿粗肽液的体外抗氧化活性[J].肉类研究, 2013, 27(6):5-9.
[20]吴宝森,谷大海,王桂瑛,等.诺邓火腿粗肽抗氧化活性及亚硝酸钠清除活性[J].肉类研究, 2017, 31(8):6-11. DOI:10.7506/rlyj1001-8123-201708002.
[21]ZHANGSiyu,ZHANGChaoyang,QIAOYan,etal.Effectof?avourzyme on proteolysis, antioxidant activity and sensory qualities ofCantonesebacon[J].FoodChemistry,2017,237:779-785.DOI:10.1016/j.foodchem.2017.06.026.
[22]LIYanhong,JIANGBo,ZHANGTao,etal.Antioxidantand freeradical-scavengingactivitiesofchickpeaproteinhydrolysate(CPH)[J].FoodChemistry,2008,106(2):444-450.DOI:10.1016/j.foodchem.2007.04.067.
[23]LEES,KIME,HWANGJ,etal.Purificationandcharacterisation ofanantioxidativepeptidefromenzymatichydrolysatesofduck processing by-products[J].Food Chemistry,2010, 123(2):216-220.DOI:10.1016/j.foodchem.2010.04.001.
[24]蔡俊,陈季旺,王茹,等.多肽体外抗氧化活性测定方法的比较[J].食品科学, 2016, 37(11):52-57. DOI:10.7506/spkx1002-6630-201611010.
[25]韦献雅,殷丽琴,钟成,等. DPPH法评价抗氧化活性研究进展[J].食品科学, 2014, 35(9):317-322. DOI:10.7506/spkx1002-6630-201409062.
[26]ZHANGW,SHANX,SAMARAWEERAH,etal.Improving functionalvalueofmeatproducts[J].MeatScience,2010,86(1):15-31. DOI:10.1016/j.meatsci.2010.04.018.
[27]SHIY,KOVACS-NOLANJ,JIANGB,etal.Antioxidantactivity of enzymatic hydrolysates from eggshell membrane proteins and its protectivecapacityinhumanintestinalepithelialCaco-2cells[J].JournalofFunctionalFoods,2014,10(3):35-45.DOI:10.1016/j.jff.2014.05.004.
[28]CACCIUTTOLO M A, TRINH L, LUMPKIN J A, et al. Hyperoxia inducesDNAdamageinmammaliancells[J].FreeRadical BiologyandMedicine,1993,14(3):267-276.DOI:10.1016/0891-5849(93)90023-N.
[29]SAIDIS,DERATANIA,BELLEVILLEMP,etal.Antioxidant propertiesofpeptidefractionsfromtunadarkmuscleprotein by-producthydrolysateproducedbymembranefractionation process[J].FoodResearchInternational,2014,65:329-336.DOI:10.1016/j.foodres.2014.09.023.
[30]KO S C, KIM D, JEON Y J. Protective effect of a novel antioxidative peptidepurifiedfromamarineChlorellaellipsoideaprotein againstfreeradical-inducedoxidativestress[J].FoodandChemical Toxicology, 2012, 50(7):2294-2302. DOI:10.1016/j.fct.2012.04.022.
[31]AIS,SOICHIT,TOSHIHIDEN.Antioxidantactivityofpeptides obtained from porcine myo?brillar proteins by protease treatment[J].Journal of Agricultural and Food Chemistry, 2003, 51(12):3661-3667.DOI:10.1021/jf021156g.
[32]荣建华,李小定,谢笔钧.大豆肽体外抗氧化效果的研究[J].食品科学, 2002, 23(11):118-120. DOI:10.3321/j.issn:1002-6630.2002.11.031.
[33]刘文颖,谷瑞增,鲁军,等.大豆低聚肽的成分分析及体外抗氧化作用[J].食品工业, 2015, 36(4):200-203.
[34]JAE-YOUNGJ,ZHONG-JIQ,SE-KWONK.Antioxidantpeptide isolated from muscle protein of bullfrog, Rana catesbeiana Shaw[J].JournalofMedicinalFood,2007,10(3):401-407.DOI:10.1089/jmf.2006.169.
[35]SAMARANAYAKA A G P, LI-CHAN E C Y. Food-derived peptidic antioxidants:a review of their production, assessment, and potential applications[J].JournalofFunctionalFoods,2011,3(4):229-254.DOI:10.1016/j.jff.2011.05.006.
[36]NAJAFIAN L, BABJI A S. Isolation, puri?cation and identi?cation ofthreenovelantioxidativepeptidesfrompatin(Pangasiussutchi)myofibrillarproteinhydrolysates[J].LWT-FoodScienceand Technology, 2015, 60(1):452-461. DOI:10.1016/j.lwt.2014.07.046.
[2]REICHMANND,VOTHW,JAKOBU.Maintainingahealthy proteome during oxidative stress[J]. Molecular Cell, 2018, 69(2):203-213.DOI:10.1016/j.molcel.2017.12.021.
[3]李兴太,张春英,仲伟利,等.活性氧的生成与健康和疾病关系研究进展[J].食品科学, 2016, 37(13):257-270. DOI:10.7506/spkx1002-6630-201613046.
[4]GüL?IN?. Antioxidant activity of food constituents:an overview[J].ArchivesofToxicology,2012,86(3):345-391.DOI:10.1016/j.peptides.2010.06.020.
[5]SARMADIBH,ISMAILA.Antioxidativepeptidesfrom foodproteins:areview[J].Peptides,2010,31(10):1949-1956.DOI:10.1016/j.peptides.2010.06.020.
[6]ZHU Chaozhi, ZHANG Wangang, KANG Zhuangli, et al. Stability of an antioxidant peptide extracted from Jinhua ham[J]. Meat Science,2014, 96(2):783-789. DOI:10.1016/j.meatsci.2013.09.004.
[7]XINGLujuan,HUYaya,HUHongyan,etal.Purificationand identificationofantioxidativepeptidesfromdry-curedXuanwei ham[J].FoodChemistry,2016,194:951-958.DOI:10.1016/j.foodchem.2015.08.101.
[8]WANG Lusha, HUANG Jichao, CHEN Yulian, et al. Identi?cation and characterization of antioxidant peptides from enzymatic hydrolysates of duck meat[J]. Journal of Agricultural and Food Chemistry, 2015,63(13):3437-3444. DOI:10.1021/jf506120w.
[9]FENG Li, QIAO Yan, ZOU Yufeng, et al. Effect of?avourzyme on proteolysis,antioxidantcapacityandsensoryattributesofChinese sausage[J].MeatScience,2014,98(1):34-40.DOI:10.1016/j.meatsci.2014.04.001.
[10]ZHU Chaozhi, ZHANG Wangang, ZHOU Guanghong, et al. Isolation andidentificationofantioxidantpeptidesfromJinhuaham[J].Journal of Agricultural and Food Chemistry, 2013, 61(6):1265-1271.DOI:10.1021/jf3044764.
[11]孔令明,李芳,陶永霞,等.多肽的功能活性与研究进展[J].中国食品添加剂,2 0 0 9,6 7(3):7 1-7 3.D O I:1 0. 3 9 6 9/j.issn.1006-2513.2009.03.015.
[12]罗雨婷,吴宝森,谷大海,等.不同加工年份诺邓火腿粗肽的特征与抗氧化活性[J].肉类研究, 2017, 31(9):32-37. DOI:10.7506/rlyj1001-8123-201709006.
[13]赵改名,周光宏,柳艳霞,等.肌肉非蛋白氮和游离氨基酸在金华火腿加工过程中的变化[J].食品科学, 2006, 27(2):33-37.DOI:10.3321/j.issn:1002-6630.2006.02.001.
[14]MORAL,BOLUMART,HERESA,etal.Effectofcookingand simulatedgastrointestinaldigestionontheactivityofgenerated bioactivepeptidesinagedbeefmeat[J].FoodandFunction,2017,8(12):4347-4355. DOI:10.1039/C7FO01148B.
[15]GALLEGOM,MORAL,HAYESM,etal.Effectofcooking andinvitrodigestionontheantioxidantactivityofdry-curedham by-products[J].FoodResearchInternational,2017,97:296-306.DOI:10.1016/j.foodres.2017.04.027.
[16]SUN Weizheng, ZHAO Haifeng, ZHAO Qiangzhong, et al. Structural characteristics of peptides extracted from Cantonese sausage during dryingandtheirantioxidantactivities[J].InnovativeFoodScience andEmergingTechnologies,2009,10(4):558-563.DOI:10.1016/j.ifset.2009.07.006.
[17]陈好春,周兴虎,康大成,等.风味蛋白酶对腌腊鸡腿蛋白质降解、抗氧化能力和感官品质的影响[J].食品与发酵工业, 2016(7):128-134. DOI:10.13995/j.cnki.11-1802/ts.201607022.
[18]杨凯,李迎楠,李享,等.基于计算机视觉技术分析猪肉对清酱肉挥发性风味成分的影响[J].肉类研究, 2018, 32(4):39-43.DOI:10.7506/rlyj1001-8123-201804007.
[19]祝超智,张万刚,徐幸莲,等.金华火腿粗肽液的体外抗氧化活性[J].肉类研究, 2013, 27(6):5-9.
[20]吴宝森,谷大海,王桂瑛,等.诺邓火腿粗肽抗氧化活性及亚硝酸钠清除活性[J].肉类研究, 2017, 31(8):6-11. DOI:10.7506/rlyj1001-8123-201708002.
[21]ZHANGSiyu,ZHANGChaoyang,QIAOYan,etal.Effectof?avourzyme on proteolysis, antioxidant activity and sensory qualities ofCantonesebacon[J].FoodChemistry,2017,237:779-785.DOI:10.1016/j.foodchem.2017.06.026.
[22]LIYanhong,JIANGBo,ZHANGTao,etal.Antioxidantand freeradical-scavengingactivitiesofchickpeaproteinhydrolysate(CPH)[J].FoodChemistry,2008,106(2):444-450.DOI:10.1016/j.foodchem.2007.04.067.
[23]LEES,KIME,HWANGJ,etal.Purificationandcharacterisation ofanantioxidativepeptidefromenzymatichydrolysatesofduck processing by-products[J].Food Chemistry,2010, 123(2):216-220.DOI:10.1016/j.foodchem.2010.04.001.
[24]蔡俊,陈季旺,王茹,等.多肽体外抗氧化活性测定方法的比较[J].食品科学, 2016, 37(11):52-57. DOI:10.7506/spkx1002-6630-201611010.
[25]韦献雅,殷丽琴,钟成,等. DPPH法评价抗氧化活性研究进展[J].食品科学, 2014, 35(9):317-322. DOI:10.7506/spkx1002-6630-201409062.
[26]ZHANGW,SHANX,SAMARAWEERAH,etal.Improving functionalvalueofmeatproducts[J].MeatScience,2010,86(1):15-31. DOI:10.1016/j.meatsci.2010.04.018.
[27]SHIY,KOVACS-NOLANJ,JIANGB,etal.Antioxidantactivity of enzymatic hydrolysates from eggshell membrane proteins and its protectivecapacityinhumanintestinalepithelialCaco-2cells[J].JournalofFunctionalFoods,2014,10(3):35-45.DOI:10.1016/j.jff.2014.05.004.
[28]CACCIUTTOLO M A, TRINH L, LUMPKIN J A, et al. Hyperoxia inducesDNAdamageinmammaliancells[J].FreeRadical BiologyandMedicine,1993,14(3):267-276.DOI:10.1016/0891-5849(93)90023-N.
[29]SAIDIS,DERATANIA,BELLEVILLEMP,etal.Antioxidant propertiesofpeptidefractionsfromtunadarkmuscleprotein by-producthydrolysateproducedbymembranefractionation process[J].FoodResearchInternational,2014,65:329-336.DOI:10.1016/j.foodres.2014.09.023.
[30]KO S C, KIM D, JEON Y J. Protective effect of a novel antioxidative peptidepurifiedfromamarineChlorellaellipsoideaprotein againstfreeradical-inducedoxidativestress[J].FoodandChemical Toxicology, 2012, 50(7):2294-2302. DOI:10.1016/j.fct.2012.04.022.
[31]AIS,SOICHIT,TOSHIHIDEN.Antioxidantactivityofpeptides obtained from porcine myo?brillar proteins by protease treatment[J].Journal of Agricultural and Food Chemistry, 2003, 51(12):3661-3667.DOI:10.1021/jf021156g.
[32]荣建华,李小定,谢笔钧.大豆肽体外抗氧化效果的研究[J].食品科学, 2002, 23(11):118-120. DOI:10.3321/j.issn:1002-6630.2002.11.031.
[33]刘文颖,谷瑞增,鲁军,等.大豆低聚肽的成分分析及体外抗氧化作用[J].食品工业, 2015, 36(4):200-203.
[34]JAE-YOUNGJ,ZHONG-JIQ,SE-KWONK.Antioxidantpeptide isolated from muscle protein of bullfrog, Rana catesbeiana Shaw[J].JournalofMedicinalFood,2007,10(3):401-407.DOI:10.1089/jmf.2006.169.
[35]SAMARANAYAKA A G P, LI-CHAN E C Y. Food-derived peptidic antioxidants:a review of their production, assessment, and potential applications[J].JournalofFunctionalFoods,2011,3(4):229-254.DOI:10.1016/j.jff.2011.05.006.
[36]NAJAFIAN L, BABJI A S. Isolation, puri?cation and identi?cation ofthreenovelantioxidativepeptidesfrompatin(Pangasiussutchi)myofibrillarproteinhydrolysates[J].LWT-FoodScienceand Technology, 2015, 60(1):452-461. DOI:10.1016/j.lwt.2014.07.046.